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Open Access Highly Accessed Technical Note

An RNA isolation system for plant tissues rich in secondary metabolites

Sanjay Ghawana12, Asosii Paul1, Hitesh Kumar13, Arun Kumar1, Harsharan Singh14, Pardeep K Bhardwaj15, Arti Rani16, Ravi S Singh1, Jyoti Raizada1, Kashmir Singh17 and Sanjay Kumar1*

Author Affiliations

1 Biotechnology Division, Institute of Himalayan Bioresource Technology (CSIR), Palampur-176 061, Himachal Pradesh, India

2 National Institute of Plant Genome Research, Aruna Asaf Ali Marg, P.O. Box No. 10531, New Delhi- 110 067, India

3 Assistant Professor, Department of Botany, SCVB Government College, Palampur-176 061, Himachal Pradesh, India

4 Assistant Professor, Biotechnology Division, Lyallpur Khalsa College, Jalandhar-144 001, Punjab, India

5 Scientist, Regional Centre of Institute of Bioresources and Sustainable Development (DBT), Tadong-737 102, Sikkim, India

6 Scientist, Vittal Mallya Scientific Research Foundation, #94/3 & 94/5, 23rd cross, 29th main, BTM II Stage, Bangalore-560 076, Karnataka, India

7 Assistant Professor, Department of Biotechnology, Panjab University, Chandigarh-160 014, India

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BMC Research Notes 2011, 4:85  doi:10.1186/1756-0500-4-85

Published: 28 March 2011

Abstract

Background

Secondary metabolites are reported to interfere with the isolation of RNA particularly with the recipes that use guanidinium-based salt. Such interference was observed in isolation of RNA with medicinal plants rheum (Rheum australe) and arnebia (Arnebia euchroma). A rapid and less cumbersome system for isolation of RNA was essential to facilitate any study related to gene expression.

Findings

An RNA isolation system free of guanidinium salt was developed that successfully isolated RNA from rheum and arnebia. The method took about 45 min and was successfully evaluated on twenty one tissues with varied secondary metabolites. The A260/280 ratio ranged between 1.8 - 2.0 with distinct 28 S and 18 S rRNA bands visible on a formaldehyde-agarose gel.

Conclusions

The present manuscript describes a rapid protocol for isolation of RNA, which works well with all the tissues examined so far. The remarkable feature was the success in isolation of RNA with those tissues, wherein the most commonly used methods failed. Isolated RNA was amenable to downstream applications such as reverse transcription-polymerase chain reaction (RT-PCR), differential display (DD), suppression subtractive hybridization (SSH) library construction, and northern hybridization.